Table 1 summarizes information on the major collections of Orthotylinae and Phylinae and their geographic strengths. Existing collections tend to be concentrated in the large North American and European museums, with the conspicuous exception of the Australian Museum. Many smaller museums may also have relevant materials. We will seek out such collections, especially through contact with members of the International Heteropterists Society.

Of particular interest to this study is material from Australia, which is only now becoming available with sufficient geographic coverage to make comprehensive study feasible, as noted above. Additionally, there is a large amount of well-documented modern material available from western North America (American Museum, Smithsonian Institution), most of it collected by team members Schuh, Henry, and Schwartz, as well as the critical collections of J. C. Schaffner (Texas A&M University) from Mexico. Although much of this material has been the subject of modern revisionary work (e.g., Schuh and Schwartz, 1985, 1988; Schuh, 2000a, b, 2001; Stonedahl, 1990; Stonedahl and Schuh, 1986; Stonedahl and Schwartz, 1986), several clades remain to be studied; it is this portion of available Nearctic material that will add a significant component of basic descriptive and revisionary work to our PBI study. Collections of the Zoological Institute, St. Petersburg, are the richest source for Palearctic material.

Although we are generally familiar with museum holdings world wide, we have budgeted for sufficient museum visits to allow for examination of existing collections by specialists on our team, to assure that all materials pertinent to this study are located and incorporated.

Additional Collecting

Table 2 gives the numbers of described and estimated undescribed species of Orthotylinae and Phylinae. These figures are based on our knowledge of the literature and collections, information provided by colleagues, and the species-level diversity estimates provided above for Australia. This summary indicates known tribal-level diversity by region and the amount of additional diversity we estimate can be documented through targeted collecting and detailed analysis of existing collections.

Table 1. PRINCIPLE WORLD
COLLECTIONS OF ORTHOTYLINAE AND PHYLINAE

Collection

Geographic Coverage

No. specimens

% studied

% Databased

No. Studied

No. databased

American Museum of Natural History

World (significant Australian
holdings)

150,000

0.50

0.25

75,000

37,500

Australian Museum, Sydney

Australia*

58,000

0.05

0.60

3,000

35,000

Bishop Museum, Honolulu

Tropical Asia, Pacific Islands

15,000

0.65

0.50

10,000

7,500

California Academy of Sciences

World (important Nearctic holdings)

15,000

0.75

0.33

12,000

5,000

Canadian National Collection

Nearctic including Mexico

50,000

0.75

0.50

37,500

25,000

Hamburg University

Palearctic (important for
reference)

6,000

0.95

0.00

5,700

0

Linnavuori Collection, Turku,
Finland

Middle East, Africa

40,000

0.90

0.00

36,000

0

Museu Nacional, Rio de Janeiro

mostly Neotropical (many types)

2,000

0.99

0.00

1,980

0

Museum d'Histoire Naturelle, Geneva

Palearctic (no figures available)

Museum d'Histoire Naturelle, Paris

Europe, Africa, Madagascar

10,000

0.65

0.00

6,500

0

Museum Zoology, LIPI, Bogor

Tropical Asia

2,000

0.00

0.00

0

0

Nankai University Insect Collection

China

5,000

3,000

0

Natuurhistorisch Museum, Leiden

Tropical Asia (no figures
available)

0.10

0.00

Natural History Museum, London

World (historical; many types)

10,000

0.95

0.00

9,500

0

Plant Protection Res. Inst.,
Pretoria

South Africa

3,000

0.50

0.00

1,500

0

Royal Central African Museum

Central Africa

2,000

0.50

0.00

1,000

0

Smithsonian Institution

World (most Nearctic types)

75,000

0.70

0.33

50,000

25,000

Texas A&M University

Mexico, SE USA

25,000

0.40

0.10

10,000

2,500

Zoological Lab., Okayama University

mostly Eastern Asia

10,000

3,000

6,000

Zoological Institute, St.
Petersburg

World (premier Palearctic
collection)

75,000

0.85

0.00

60,000

0

Zoological Museum, Helsiniki

World (historical; some types)

2,000

0.99

0.00

1,980

0

Totals

555,000

0.26

327,660

143,500

*other Australian collections on
loan included

These data suggest that 73% of the species have been described and that 89% of the undescribed species diversity exists in the Southern Hemisphere. On the basis of this evidence, we propose to conduct the fieldwork regime described in Table 3, with the resources devoted to the Southern Hemisphere being roughly proportional to the degree the fauna requires further study. The rationale for our collecting focus is influenced by the need to acquire as much new host-documented material as possible. Our field experience suggests the greatest return for effort expended will come from working in the subtropical and temperate areas of the world.

Table 2. DESCRIBED AND ESTIMATED NUMBERS OF SPECIES
BY TRIBE AND REGION

(predicted additional species in parentheses; see
text for explanation of methods of prediction)

Phylinae

Orthotylinae

Totals

Region/ Tribe

Auricillocorini

Hallodapini

Leucophoropterini

Phylini

Pilophorini

Halticini

Nichomachini

Orthotylini

Total Described Species

Total Predicted Species

Grand Total

Australian

1

40 (90)

8 (400)

(15)

(10)

30 (450)

79

955

1024

Ethiopian

104 (15)

10 (15)

176 (75)

36

12

11

82 (50)

431

155

586

Nearctic

52 (15)

1

405 (70)

43

18

507 (50)

1026

135

1161

Neotropical

2

6

121

4

3

378 (50)

514

50

564

Oriental

14 (10)

25

58 (15)

147 (50)

37

15(10)

166 (50)

462

115

577

Palearctic

70

5

846

35

161

4

272 (10)

1393

10

1403

Described Species

14

254

120

1703

155

209

15

1435

3905

Predicted Species

10

30

120

595

15

20

660

1450

Total Species

24

284

240

2298

170

229

15

2095

5355

Table 3. PROPOSED SCHEDULE OF FIELD WORK

Region/Area

No. visits

Field Work Coordinator

Team Size

Project year/quarter

Duration in days

Estimated Cost

Tasmania

1

Cassis

3

1/2

21

$10,245

Mexico

1

Schwartz

3

1/3

15

$7,350

South Africa, Eastern Cape, Karoo

1

Schuh

3

1/4

21

$9,750

Southern Brazil

1

Henry

3

1/3

15

$7,500

$34,845

Central Queensland

1

Cassis

3

2/1

21

$10,245

New Caledonia, Vanuatu

1

Cassis

3

2/3

21

$10,245

SE Asia

1

Yasunaga

3

2/2

21

$10,875

South Africa, Namaqualand

1

Schuh

3

2/1

21

$9,750

Southern China

1

Yasunaga

3

2/3

15

$6,900

Western United States

1

Schwartz

3

2/3

15

$7,350

$55,365

Argentina/Chile*

1

Henry

3

3/1

15

$7,500

Geraldton-Pilbara

1

Cassis

3

3/3

21

$10,245

SE Asia

1

Yasunaga

3

3/2

21

$10,875

South Africa, Western Cape

1

Schuh

3

3/1

21

$9,750

Southern China

1

Yasunaga

3

3/3

15

$6,900

$45,270

Totals

15

279

$135,480

$135,480

* PI Schuh collected widely in Chile & Argentina in 1981, 1986, 1993. His collections will be part of this PBI study.

Although collecting in the tropics will undoubtedly yield additional taxa, the probability of substantially increasing knowledge, particularly of host associations, is small compared to effort required. Thus, our fieldwork in the tropics is targeted at acquiring generic representation, particularly for acquisition of DNA sequence data. In contrast, our efforts in temperate and subtropical efforts, the latitudes where the Orthotylinae and Phylinae show their greatest diversity, will be systematic in planning and coverage, focusing on taxonomic and geographic gaps.

Several as yet unmentioned geographic areas of importance to biodiversity and biogeography in the Orthotylinae and Phylinae require comment. New Zealand has a small Phylinae fauna (Eyles and Schuh, in press); the equally small fauna of Orthotylinae awaits study. For Madagascar we will seek out existing material, particularly in Paris, collections made with BS&I support by the Center for Biodiversity and Conservation at the American Museum of Natural History (AMNH), and the program of the California Academy of Sciences currently supported by the NSF. The Phylinae faunas of India and Sri Lanka were dealt with by Schuh (1984), incorporating the collections made by the Lund University Expedition, among others. The Orthotylinae remain to be studied. Senior investigator T. J. Henry has conducted field work in Sri Lanka and has contacts there. We will bring knowledge of the Orthotylinae to the same level that exists for the Phylinae. The largest clades in the speciose orthotyline fauna of Hawaii have been revised recently (Sarona Kirkaldy [Asquith, 1994]; Nesiomiris Kirkaldy [Gagne, 1997]; Orthotylus Fieber [Polhemus, in press]).

Fieldwork will be conducted in the first three years of the project. This will allow new specimen materials to be included in our final published and web-based products. To achieve this end, all specimen materials should be mounted and labeled early in the fourth grant year.

Specimen Sampling and Data Collection Protocols

The following procedures were perfected through our own field and laboratory experience.

Techniques of Collection and Preparation

Collection and Preservation of Dried Specimens: Members of the Orthotylinae and Phylinae are frequently associated with new growth and flowers (Kullenberg, 1944; Wheeler, 2001). Thus, locating specimens is strongly correlated with the discovery of plants in flower or with developing foliage. Shallow oval nets are employed because they allow for easy collection of large numbers of bugs and determining from which plant, or plant part, the bugs came.

Specimens are killed in the field, stored dry, and transported back to the lab for mounting.

Preservation of Specimens for DNA Sequencing: We will take every opportunity to preserve specimens for DNA sequencing. Emphasis will be on sampling as many genera across all tribes as possible, to enhance the sequence character database assembled by W. C. Wheeler and PI Schuh (see Results of Prior Support). Up to this time we have preserved specimens in 95-100% ethanol. The DNA of Miridae so preserved has proved remarkably easy to isolate, amplify, and sequence. Where applicable and possible we will use alternative approaches, including liquid nitrogen and buffers such as "RNA Later" that allow for the preservation of more potential information. Tissues and purified DNA from this study will be deposited in the liquid-nitrogen based Ambrose Monell Collection for Molecular and Microbial Research at the AMNH. Sequences will be submitted to GenBank upon project completion.

Host-Plant Voucher Collection, Identification, Imaging, and Databasing: Host plant voucher specimens are collected and submitted to botanical specialists for identification. The costs for plant identification in our budget are based on our experience working with the Royal Botanical Gardens, Sydney, and the Western Australian Herbarium, Perth. Digital images of vouchered plants are made using a high-resolution flat-bed scanner and included in our locality database. We capture digital images of living plants and habitats for inclusion in the locality database.

Locality-Data Acquisition in the Field: Use of GPS devices is a routine aspect of field data collection for all of our senior investigators. PIs Cassis and Schuh have successfully used a serial GPS device and recorded data directly to a laptop-based database. Data may alterna-tively be recorded as way points on handheld devices and downloaded later. All data collected in the field will be submitted to the locality database described in the Management Plan.

Preparation and Labeling of Dried Insect Specimens: Specimens will be mounted by "preparators" hired specifically for this project. Label copy is generated directly from the locality database. Labels are printed on archival quality paper, using offset printing to assure long-term stability. An "evaluation of effort" is presented in the Budget Justification.

Long Term Preservation, Curation, and Vouchering of Insect Specimens: See Management Plan for details.

Retrospective Data Capture and Geocoding

Size of the Task: As indicated in Table 1, ~400,000 specimens will need to be databased. An "evaluation of effort" for data entry is presented in the Budget Justification.

Data Capture: The format for retrospective specimen data will be consistent with that for newly acquired material. Because databasing of pre-existing collections will occur in numerous institutions with variable infrastructure, such data will be captured on laptops using a stand-alone application and later uploaded to the master database. Data quality will be controlled through a structured input interface and through review by a project "data monitor" before uploading to the master database.

Geocoding: Most retrospective data requires geocoding to allow for web mapping. We will use various methods depending on their effectiveness, including among others: looking up individual localities on a map; use of an electronic gazeteer; or, use of specialized software such as E-GAZ prepared by the Australian Biological Resources Study. We will assign "accuracy codes" to distinguish between GPS recorded data and less precisely characterized label.

Unique Specimen Identification/Bar Coding: Because the majority of databased material in the Orthotylinae and Phylinae will be processed during this project, the opportunity exists to provide unique specimen identifiers. We will implement the use of bar codes in our existing locality database, relying on experience of investigators who have employed them with pinned insects at institutions such as the University of Kansas and InBio, Costa Rica.

Judging Completeness of Sample: Within the Orthotylinae and Phylinae some species are rare in collections and others are very common. All possible specimens of "rare" species will be databased. Databasing all specimens of extremely common species makes little sense in terms of additional information added compared to effort expended. The following criteria will be used to determine how many specimens will be databased. Data entry personnel should continue to add data: 1) if range of distribution is being extended; 2) if new host data are being recorded; 3) if both sexes are not represented even if the criteria above are satisfied.